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Illumina 测序 16S rRNA 标签显示与在巴基斯坦不同生境中生长的绿豆(Vigna radiata L.)根瘤相关的根瘤菌和非根瘤菌多样性存在差异。

Illumina sequencing of 16S rRNA tag shows disparity in rhizobial and non-rhizobial diversity associated with root nodules of mung bean (Vigna radiata L.) growing in different habitats in Pakistan.

机构信息

National Institute for Biotechnology and Genetic Engineering (NIBGE), P.O. Box 577, Faisalabad, Pakistan; Pakistan Institute of Engineering and Applied Sciences (PIEAS), Nilore, Islamabad, Pakistan.

Missouri State University, Biology Department, Springfield, MO 65897, USA.

出版信息

Microbiol Res. 2020 Jan;231:126356. doi: 10.1016/j.micres.2019.126356. Epub 2019 Oct 23.

Abstract

In Rhizobium-legume symbiosis, the nodule is the most frequently studied compartment, where the endophytic/symbiotic microbiota demands critical investigation for development of specific inocula. We identified the bacterial diversity within root nodules of mung bean from different growing areas of Pakistan using Illumina sequencing of 16S rRNA gene. We observed specific OTUs related to specific site where Bradyrhizobium was found to be the dominant genus comprising of 82-94% of total rhizobia in nodules with very minor fraction of sequences from other rhizobia at three sites. In contrast, Ensifer (Sinorhizobium) was single dominant genus comprising 99.9% of total rhizobial sequences at site four. Among non-rhizobial sequences, the genus Acinetobacter was abundant (7-18% of total sequences), particularly in Bradyrhizobium-dominated nodule samples. Rhizobia and non-rhizobial PGPR isolated from nodule samples include Ensifer, Bradyrhizobium, Acinetobacter, Microbacterium and Pseudomonas strains. Co-inoculation of multi-trait PGPR Acinetobacter sp. VrB1 with either of the two rhizobia in field exhibited more positive effect on nodulation and plant growth than single-strain inoculation which favors the use of Acinetobacter as an essential component for development of mung bean inoculum. Furthermore, site-specific dominance of rhizobia and non-rhizobia revealed in this study may contribute towards decision making for development and application of specific inocula in different habitats.

摘要

在根瘤菌-豆科植物共生中,根瘤是最常被研究的器官,其中内生/共生微生物群落的需求需要进行深入研究,以开发特定的接种剂。我们使用 Illumina 16S rRNA 基因测序技术,对来自巴基斯坦不同种植区的绿豆根瘤中的细菌多样性进行了鉴定。我们观察到了与特定部位相关的特定 OTUs,其中 Bradyrhizobium 被发现是优势属,占根瘤中总根瘤菌的 82-94%,而其他根瘤菌的序列比例非常小,在三个地点的三个地点。相比之下,Ensifer(Sinorhizobium)是单一优势属,占总根瘤菌序列的 99.9%。在非根瘤菌序列中,不动杆菌属丰度较高(占总序列的 7-18%),特别是在 Bradyrhizobium 主导的根瘤样本中。从根瘤样本中分离出的根瘤菌和非根瘤促生菌(PGPR)包括 Ensifer、Bradyrhizobium、Acinetobacter、Microbacterium 和 Pseudomonas 菌株。将多性状 PGPR Acinetobacter sp. VrB1 与两种根瘤菌中的一种共接种到田间,对结瘤和植物生长的影响比单菌株接种更为积极,这有利于将不动杆菌作为绿豆接种剂开发的必要组成部分。此外,本研究中揭示的根瘤菌和非根瘤菌的特定部位优势可能有助于在不同生境中开发和应用特定接种剂的决策。

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